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World J Gastrointest Oncol. Apr 15, 2022; 14(4): 872-886
Published online Apr 15, 2022. doi: 10.4251/wjgo.v14.i4.872
Xihuang pills induce apoptosis in hepatocellular carcinoma by suppressing phosphoinositide 3-kinase/protein kinase-B/mechanistic target of rapamycin pathway
Yong-Jie Teng, Zhe Deng, Zhao-Guang Ouyang, Qing Zhou, Si Mei, Xing-Xing Fan, Yong-Rong Wu, Hong-Ping Long, Le-Yao Fang, Dong-Liang Yin, Bo-Yu Zhang, Yin-Mei Guo, Wen-Hao Zhu, Zhen Huang, Piao Zheng, Di-Min Ning, Xue-Fei Tian
Yong-Jie Teng, Qing Zhou, Hong-Ping Long, Le-Yao Fang, The First Hospital of Hunan University of Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Zhe Deng, Yong-Rong Wu, Yin-Mei Guo, Wen-Hao Zhu, Zhen Huang, Piao Zheng, Di-Min Ning, Xue-Fei Tian, College of Integrated Chinese and Western Medicine, Hunan Key Laboratory of Translational Research in Formulas and Zheng of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Zhao-Guang Ouyang, Department of Preventive Dentistry, Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine, Guangzhou Medical University, Guangzhou 510132, Guangdong Province, China
Si Mei, Department of Physiology, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Xing-Xing Fan, State Key Laboratory of Quality Research in Chinese Medicine, Macau Institute For Applied Research in Medicine and Health, Macau University of Science and Technology, Macau, China
Dong-Liang Yin, School of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Bo-Yu Zhang, College of Acupuncture and Massage, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Author contributions: Teng YJ, Deng Z, and Wu YR completed the experiment and wrote the paper; Teng YJ, Deng Z, and Ouyang ZG reviewed the manuscript; Tian XF and Zhou Q provided research ideas; Long HP and Fang LY performed HPLC analysis; Zhang BY has processed the figures of the article; Fan XX, Yin DL, Guo YM, Zheng P, Huang Z and Ning DM conducted statistical analysis; all authors have read and approved the manuscript to ensure its accuracy and completeness.
Supported by National Natural Science Foundation of China, No. U20A20408 and No. 82074450; Natural Science Foundation of Hunan Province, No. 2020JJ4066; Hunan Province "Domestic First-class Cultivation Discipline" Integrated Traditional Chinese and Western Medicine Open Fund Project, No. 2020ZXYJH34 and No. 2020ZXYJH35; Hunan Graduate Scientific Research Innovation Project, No. QL20210173 and No. CX20210730; Hunan Province Science and Technology Innovation Talents Plan College Students Science and Technology Innovation and Entrepreneurship Project, No. 2020RC1004; Guangzhou Health Science and Technology Project, No. 20221A011102; and Hunan Traditional Chinese Medicine Scientific Research Project, No. 202101.
Institutional animal care and use committee statement: This study was reviewed and approved by the Ethics Review Committee of Experimental Animal Welfare at the Central South University in Changsha, China.
Conflict-of-interest statement: The authors declare that they have no competing interests.
Data sharing statement: Dataset available from the corresponding author at
003640@hnucm.edu.cn. Participants gave informed consent for data sharing.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Xue-Fei Tian, PhD, Professor, College of Integrated Chinese and Western Medicine, Hunan Key Laboratory of Translational Research in Formulas and Zheng of Traditional Chinese Medicine, Hunan University of Chinese Medicine, No. 300 Xueshi Road, Yuelu District, Changsha 410208, Hunan Province, China.
003640@hnucm.edu.cn
Received: September 7, 2021
Peer-review started: September 7, 2021
First decision: December 4, 2021
Revised: December 30, 2021
Accepted: March 14, 2022
Article in press: March 14, 2022
Published online: April 15, 2022
Processing time: 219 Days and 19 Hours
ARTICLE HIGHLIGHTS
Research background
Xihuang pills (XHP) are a traditional Chinese preparation with antitumour properties. They inhibit the growth of breast cancer, glioma, and other tumours by regulating the phosphoinositide 3-kinase/protein kinase-B/mechanistic target of rapamycin (PI3K/Akt/mTOR) signalling pathway. However, the effects and mechanisms of action of XHP in hepatocellular carcinoma (HCC) remain unclear. Regulation of the PI3K/Akt/mTOR signalling pathway effectively inhibits the progression of HCC.
Research motivation
We hypothesized that XHP might play a role in inhibiting HCC through the PI3K/Akt/mTOR signalling pathway.
Research objectives
To confirm the effect of XHP on HCC and the possible mechanisms involved.
Research methods
The chemical constituents and active components of XHP were analysed using ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry (MS) (UPLC-Q-TOF-MS). First, cell-based experiments and in vivo xenograft tumour experiments were utilized to evaluate the effect of XHP on HCC tumorigenesis. Cell viability was assessed using the CCK-8 assay, followed by an assessment of cell migration using a wound healing assay. Second, the effect of XHP on the apoptosis of SMMC-7721 cells was evaluated. Third, Western blotting and reverse-transcription polymerase chain reaction were performed to confirm the effects of XHP on the protein and mRNA expression of components of the PI3K/Akt/mTOR signalling pathway. Finally, the effects of XHP on the tumorigenesis of subcutaneous hepatocellular tumours in nude mice were assessed.
Research results
The 12 compounds were identified in XHP by high-resolution MS. The cell viability assay results showed that treatment with 0.625 mg/mL XHP extract decreased HCC cell viability after 12 h. Moreover, XHP significantly inhibited cell migration and resulted in cell cycle arrest and apoptosis. Furthermore, XHP downregulated the PI3K/Akt/mTOR signalling pathway, which activated apoptosis executioner proteins (e.g., caspase-9 and caspase-3). The inhibitory effects of XHP on HCC cell growth were determined in vivo by analysing the tumour xenograft volumes and weights.
Research conclusions
XHP inhibited HCC cell growth and migration by stimulating apoptosis via the downregulation of the PI3K/Akt/mTOR signalling pathway, followed by the activation of caspase-9 and caspase-3. Our findings clarified that the antitumour effects of XHP on HCC cells are mediated by the PI3K/Akt/mTOR signalling pathway.
Research perspectives
Our findings revealed that XHP may be a potential complementary therapy for HCC.
